Methylation profiling using methylated DNA immunoprecipitation and tiling array hybridization.
Methylation profiling using methylated DNA immunoprecipitation and tiling array hybridization.
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DOI：
10.1007/978-1-61779-436-0_10
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Chan, Wai-Yee
中科院分区：
文献类型：
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作者：
Cheung, Hoi-Hung;Lee, Tin-Lap;Rennert, Owen M;Chan, Wai-Yee
DNA甲基化是一种重要的表观遗传修饰，调节发育，在许多疾病的病理生理学中发挥作用。它在生殖系发育过程中是动态变化的。甲基化DNA免疫沉淀法(MeDIP)是一种高效、经济的位点特异性和全基因组分析方法。甲基化的DNA片段被5-甲基胞苷识别抗体富集，因此可以同时分析CpG和非CpG甲基化。丰富的DNA片段可以被扩增并杂交成覆盖CpG岛、启动子或整个基因组的平铺阵列。通过比较不同的甲基组，可以发现在疾病或组织特异性表达中可能重要的差异甲基化区域。在这里，我们描述了一种已建立的MeDIP协议和平铺阵列杂交方法，用于分析睾丸生殖细胞的甲基化。
DNA methylation is an important epigenetic modification that regulates development and plays a role in the pathophysiology of many diseases. It is dynamically changed during germline development. Methylated DNA immunoprecipitation (MeDIP) is an efficient, cost-effective method for locus-specific and genome-wide analysis. Methylated DNA fragments are enriched by a 5-methylcytidine-recognizing antibody, therefore allowing the analysis of both CpG and non-CpG methylation. The enriched DNA fragments can be amplified and hybridized to tiling arrays covering CpG islands, promoters, or the entire genome. Comparison of different methylomes permits the discovery of differentially methylated regions that might be important in disease- or tissue-specific expression. Here, we describe an established MeDIP protocol and tiling array hybridization method for profiling methylation of testicular germ cells.